Depletion of intracellular Ca2+ stores, mediated by Mg2+-stimulated InsP3 liberation or thapsigargin, induces a capacitative Ca2+ influx in prawn oocytes.
By voltage clamp technique and intracellular calcium measurements, we recorded in prawn oocytes simultaneous [Ca2+]i and ionic current changes stimulated by external Mg2+. The [Ca2+]i response consists of an oscillation period followed by a second state of sustained [Ca2+]i level. The oscillation period successively comprises a first [Ca2+]i peak, a series of [Ca2+]i transients, and a [Ca2+]i oscillatory plateau respectively concurrent with an initial transient outward K+Ca current, an inward Na+Ca current, and a final K+Ca outward current. By using inhibitor (heparin) or sensitizers (thimerosal or caffeine) of calcium release ER channels, and caged InsP3, we established that InsP3 is the sole second messenger releasing Ca2+ from intracellular stores. By sequential substitutions and reapplications of external Ca2+, and using econazole (50 microM), a Ca2+ influx inhibitor, we documented Ca2+ influx during the [Ca2+]i oscillatory plateau. The intracellular Ca2+ store was depleted with thapsigargin (75-350 nM) in Ca2+-free ASW. Reapplication of external Ca2+ evoked a rise in [Ca2+]i, indicating a store-dependent capacitative Ca2+ influx, correlated with a K+Ca outward current increase. No measurable Ca2+ release-activated Ca2+ current (Icrac) could be detected, but was indirectly demonstrated using the sensitivity of the K+Ca channels to [Ca2+]i. We propose that the involvement of external Ca2+, in the physiological [Ca2+]i response of prawn oocytes to external Mg2+, consists of a store-dependent capacitative Ca2+ influx.
Goudeau
H
H
Observatoire Océanographique et de Biologie marine de Roscoff, UPR CNRS 9042, France. hgoudeau@sbr.sb-roscoff.fr
Goudeau
M
M
D000818:Animals; D002110:Caffeine / Q000494:pharmacology; D002118:Calcium / Q000037:antagonists & inhibitors / Q000378:metabolism* / Q000502:physiology*; D015220:Calcium Channels / Q000378:metabolism; D002462:Cell Membrane / Q000187:drug effects / Q000502:physiology; D033364:Decapoda; D005260:Female; D006493:Heparin / Q000494:pharmacology; D015544:Inositol 1,4,5-Trisphosphate / Q000031:analogs & derivatives / Q000378:metabolism* / Q000502:physiology; D007424:Intracellular Fluid / Q000378:metabolism*; D008274:Magnesium / Q000494:pharmacology*; D008845:Microinjections; D009865:Oocytes / Q000187:drug effects / Q000378:metabolism*; D018408:Patch-Clamp Techniques; D010782:Photolysis; D015221:Potassium Channels / Q000378:metabolism; D019284:Thapsigargin / Q000494:pharmacology*; D013849:Thimerosal / Q000494:pharmacology
D015220:Calcium Channels; D015221:Potassium Channels; C061868:inositol 1,4,5-trisphosphate 1-(2-nitrophenyl)ethyl ester; D013849:Thimerosal; D002110:Caffeine; D019284:Thapsigargin; D015544:Inositol 1,4,5-Trisphosphate; D006493:Heparin; D008274:Magnesium; D002118:Calcium